Analysis of monomeric Aβ (1-40) peptide by capillary electrophoresisElectronic supplementary information (ESI) available: Electropherograms from Fig. 2 plotted vs. time. See DOI: 10.1039/c0an00080a

• Main Authors: Picou, Ryan, Moses, Julia P, Wellman, Amber D, Kheterpal, Indu, Gilman, S. Douglass
• Format: Article
• Language: English
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c0an00080a

A method was developed to characterize and quantify preparations of monomeric β-amyloid (Aβ) peptide using capillary electrophoresis (CE) with UV absorbance detection. The detection limit for Aβ monomer using this method was 0.5 µM (19 pg). The self-assembly of Aβ to form amyloid fibrils is closely linked to Alzheimer's disease and is the subject of intense investigations. Consistent preparation of Aβ monomer samples at known concentrations and free of aggregates is a significant challenge for researchers studying the mechanism of Aβ fibril formation and searching for small molecules that inhibit Aβ fibril formation. Samples of Aβ monomer are known to sometimes contain pre-existing aggregates that can affect the kinetics and structure of amyloid fibrils. The CE method presented here showed that some of the monomeric Aβ samples prepared for this study contained a species producing a second peak (in addition to the major monomer peak). The aggregation was monitored using a thioflavin T fluorescence assay, and the resulting fibrils were characterized by transmission electron microscopy. Monomer samples containing the additional peak based on CE analysis were shown to aggregate more rapidly than monomer samples that were free of this putative Aβ aggregate peak. Capillary electrophoresis indicates the presence of unwanted aggregates that significantly alter the aggregation kinetics of β-amyloid peptide monomer preparations.